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Nová HPLC metoda pro stanovení nafazolinu v očních přípravcích
Dulavová, Martina ; Chocholoušová Havlíková, Lucie (advisor) ; Matysová, Ludmila (referee)
Charles University in Prague, Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate: Martina Dulavová Supervisor: PharmDr. Lucie Havlíková, Ph.D., Ass.Prof. Hannelore Kopelent Title of Diploma Thesis: A New Selective and Stability Indicating HPLC Assay for the Determination of Naphazoline in Preparations for Ocular Use Naphazoline is a 2-imidazolidine derivated drug and alpha-adrenergic agonist with vasoconstrictive and decongestive properties. Naphazoline is indicated for the therapy of rhinitis, sinusitis or allergic conjunctivitis. Naphazoline is used in liquid formulations for ophthalmic and nasal application. Naphazoline is marketed in a number of commercially available products; for example, it is contained in Coldan® Augentropfen. Due to economic and therapeutic reasons, hospital pharmacies produce miscellaneous in-house preparations. For our study, two different preparations manufactured in the sterile production of a hospital pharmacy containing naphazoline are investigated. The first formulation is based on two commercially available products and it is a mixture of Coldan® Augentropfen and Okuzell® Augentropfen at a ratio of 1:9. The second formulation is Bor-Naphazolin Augentropfen and it is completely prepared in a hospital pharmacy. In addition to...
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Development of GC Metod for the Determination of Vitamine E Acetate in Nutritive Supplements
Kymlová, Helena ; Šatínský, Dalibor (advisor) ; Chocholoušová Havlíková, Lucie (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate: Mgr. Helena Kymlová Consultant: Doc. RNDr. Dalibor Šatínský, PhD. Title of Thesis: Development of GC metod for the determination of vitamine E acetate in nutritive supplements A GC method was developed for the determination of content of vitamine E acetate in nutritive supplements. The method was optimised and validated. The column Capillary GC, Equity Columns, Equity TM - 5 poly (5 % bifenyl / 95 % dimethylpolysiloxan); 0,32 mm ID x 30 m; 1,0 mm dr was used for the analysis. Helium was used as a carrier. Split was 1 : 100. Flame-ionization detector was used for detection. Optimal conditions for analysis: temperature of column 340 řC, temperature of injection 340 řC, temperature of detector 320 řC, flow-rate 2 ml/min. Fenoxycarb was chosen as an internal standard. The method was used for an analysis of vitamine E acetate in nutritive supplements Geladrink Forte pulverized drink - pineapple and Chondrotin MSM 2600. Determined concentration of vitamine E acetate was re-counted to the content of vitamine E declared by producer.
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Development and validation of method for determination of clotrimazole in cream using HPLC
Poláčková, Jitka ; Matysová, Ludmila (advisor) ; Chocholoušová Havlíková, Lucie (referee)
The topic of my work was to optimalize and to validate the method for determination of capacity of the Clotrimazol, degradeted product (2-chlorophenyl) diphenylmethanol and conservations (methylparaben and propylparaben) in the preparation Clotrimazol ointment. My effort for this work was to develop the method that could half for total separation the particular defined substance in this sample, in the acceptable time, to find inside standard, to optimalize the conditions of separation and the method verify. The optimum chromatographic conditions were found on column Zorbax SB - Phenyl, 3,5 μm, 4,6 x 75 mm in the flow of mobile phase 0,5 ml per minute and wave - length 210 nm. Composition of the mobile phase was acetonitrile - water (55:45), pH water component was modified by acid phosphoric 5 % on 3,2. If these conditions are used, this method is validating. The testing parameters were accuracy, correctness, linearity, selectivity, robustness, detection and quantitative limit and test if the chromatographic system is acceptable.
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HPLC Determination of Lutein, Zeaxanthine and Betacarotene in Dietary Products
Dvořáková, Petra ; Šatínský, Dalibor (advisor) ; Chocholoušová Havlíková, Lucie (referee)
This rigorous thesis deals with modification of method for separation carotenoids lutein, betacarotene and zeaxanthin borrowed from diploma thesis (Optimization of chromatographic conditions for HPLC determination of chosen carotenoids, Petra Dvořáková, 2009). HPLC with VIS detection is the basic method. This method was modified by gradient elution for compressed time of analysis. Four ways of gradient elution were tested during optimization of new procedure. The second aim of this rigorous thesis is summary of extraction ways for carotenoids published in special literature and proposition of extraction procedures for carotenoids from dietary products. Extraction with organic solvents was used during searching for optimal way for extraction. Simple extraction with organic solvent was tested as basic method, but extraction with previous modification of sample (saponification or acidification of extraction medium) was tested too. In spite of testing of many various ways for extraction and many various organic solvents, there wasn't find optimal and universal extraction way.
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Determination of neopterin and its derivatives by HPLC with fluorescence detection.
Říhová, Lenka ; Chocholoušová Havlíková, Lucie (referee) ; Nováková, Lucie (advisor)
TITLE: Determination of neopterin and its derivates by HPLC with fluorescence detection SUMMARY: Neopterin, a pteridine derivative, is produced in human organism by monocytes or by B - lymphocytes stimulated with interferon γ or interleukine - 2. These active substances (interferon γ, interleukine - 2) are connected with immune system activation. Neopterin measurement in body fluids can therefore serve for monitoring of activation of various components of immune system induced by cytokines. Immune activation occurs for example during many infectious diseases, autoimmune diseases and malignant diseases. Recently the connection between neopterin and 7,8 - dihydroneopterin and intracellular oxidative stress and cell apoptosis was demonstrated in several scientific studies. Values of pteridines probably correlate with certain neurological disesases. Totally reduced form of biopterin serves as cofactor of some enzymes in human organism. In clinical practice the concentration of neopterin, eventually other related substances is expressed as the ratio neopterin / creatinine. This diploma thesis is dealing with the development of an analytical method for the determination of biologically active substances biopterin, 7,8 - dihydroneopterin, 5,6,7,8 - tetrahydroneopterin, neopterin and creatinine by high performance...
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The comparison of particle stationary phases with monolith in analysis of selected medicaments
Šestakova, Veronika ; Chocholoušová Havlíková, Lucie (referee) ; Matysová, Ludmila (advisor)
The performance of monolithic HPLC columns Chromolith® (made by Merck, Germany) and Onyx™ (made by Phenomenex, USA) and conventional C18 columns Discovery® (made by Supelco, Sigma-Aldrich, Prague, Czech Republic) and LiChroCart® 125-4 Purospher 5 µm (made by Merck, Germany) was tested and the comparison for two topical preparation Estrogel gel and Aphlox-K paste was made. The composition of mobile phase - for Estrogel gel analysis a mixture of acetonitrile, methanol and water (23:24:53, v/v/v) and for Aphlox-K paste analysis a mixture of methanol, water and acetic acid (50:50:1, v/v/v) - was usually not optimal for analysis at all types of columns. Thus an adjustment of components ratio was necessary for sufficient resolution of the analyzed compounds. Various flow rates (0.3 - 5.0 ml/min) and mobile phase composition (usually increasing ratio of water content) were applied. Determination of active substances, preservatives and impurities and comparison of retention times and system suitability test parameters was accomplished. For Estrogel gel analysis, following chromatographic conditions were found: using Chromolith® Flash RP18-e (25 mm x 4.6 mm), monolith column mobile phase was acetonitrile, methanol, water (16:24:60 v/v/v) and flow-rate 2.0 ml/min. Using monolith column Onyx™ Monolithic C18-e (50 mm x...
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Application of HPLC in pharmaceutical analysis.
Šestakova, Veronika ; Chocholoušová Havlíková, Lucie (referee) ; Matysová, Ludmila (advisor)
Determination of hexachlorophene in the product SEPTONEX ointment to the Department of Analytical Chemistry performed by HPLC. This method has many advantages over previously used titrate determination. The conditions of analysis are as follows: mobile phase is composed of 85% methanol with the addition of 1% acetic acid, column Purospher Merck RP C18, 5 mm (12.5 cm x 4 mm), flow rate of the mobile phase is 1.5 ml / min, sample injection volume is 20 µl. The aim was to optimize the use of methods, i.e. find the column using the modern trends in HPLC, which would allow obtaining precise, rapid and well reproducible results under similar chromatographic conditions. Based on the literature research were selected six different analytical columns with more or less similar characteristics as the column still used. In each of these columns have been measured solutions of standards hexachlorophene and solutions of ointment SEPTONEX samples at different flow rate of mobile phase. Other conditions of analysis have been retained. From the experiment showed that the tested columns is the most appropriate column Chromolith Performance RP18-e (10 cm x 4,6 mm) at a flow rate of mobile phase 2.0 ml / min. Another challenge was to find a suitable internal standard for faster and more accurate determination of...
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Determination of vitamine C and dehydroascorbic acid using UHPLC-MS method
Proroková, Zuzana ; Chocholoušová Havlíková, Lucie (referee) ; Nováková, Lucie (advisor)
A coupling of Ultra High Performance liquid chromatography with mass spectrometry provides a technique, which is rapid and sensitive. This thesis is focused on the use of UHPLC-MS for the determination of ascorbic acid (AA) and dehydroascorbic acid (DHA). AA is a small polar molecule that acts as an antioxidant. After oxidation AA creates DHA. AA/DHA ratio is an indicator of a redox state of organism. For the determination of AA and DHA several methods have been developed, which usually do not allow the simultaneous analysis, but require multistep subtraction procedure. The optimization of UHPLC-MS method for the determination of AA and DHA include the choice of mobile and stationary phase and mass spectrometry detector set- up. The choice of appropriate conditions depended mainly on retention time and the detector response. The effect of stationary phase, concentration, pH, composition of mobile phase on retention of AA and DHA was observed. Effect of mobile phase on stability of AA and DHA was observed as well. BEH Shield RP C18, BEH HILIC and BEH Amide column were compared. The best results were achieved on column BEH Shield RP C18. Measurements with 0.1%, 0.05%, 0.01% formic acid, 0.1%, 0.05%, 0.01% acetic acid, ammonium formate at pH 3.5 and amonium acetate at pH 4.4 and 6.8 as a water...
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